Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain XL1-Blue | Proteus mirabilis |
Protein Variants | Comment | Organism |
---|---|---|
F113A | site-directed mutagenesis of the H-site residue, the mutant shows altered substrate specificity and activity compared to the wild-type enzyme | Proteus mirabilis |
G8A | site-directed mutagenesis of the H-site residue | Proteus mirabilis |
H167A | site-directed mutagenesis of the H-site residue, the mutant shows altered substrate specificity and activity compared to the wild-type enzyme | Proteus mirabilis |
additional information | mutations at the H-site can be used to modulate activity at the level of the different catalytic mechanisms operating on the chosen substrates, each mutation showing a different fingerprint, overview | Proteus mirabilis |
S110A | site-directed mutagenesis of the H-site residue, the mutant shows altered substrate specificity and activity compared to the wild-type enzyme | Proteus mirabilis |
W164A | site-directed mutagenesis of the H-site residue the mutant shows altered substrate specificity and activity compared to the wild-type enzyme | Proteus mirabilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.453 | - |
1-Fluoro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant W164A | Proteus mirabilis | |
0.557 | - |
glutathione | pH 6.5, 25°C, mutant H167A | Proteus mirabilis | |
0.577 | - |
1-Fluoro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant F113A | Proteus mirabilis | |
0.599 | - |
1-Fluoro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant H167A | Proteus mirabilis | |
0.651 | - |
glutathione | pH 6.5, 25°C, wild-type enzyme | Proteus mirabilis | |
0.653 | - |
glutathione | pH 6.5, 25°C, mutant S110A | Proteus mirabilis | |
0.69 | - |
glutathione | pH 6.5, 25°C, mutant H167A | Proteus mirabilis | |
0.696 | - |
glutathione | pH 6.5, 25°C, mutant S110A | Proteus mirabilis | |
0.732 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant W164A | Proteus mirabilis | |
0.76 | - |
1-Fluoro-2,4-dinitrobenzene | pH 6.5, 25°C, wild-type enzyme | Proteus mirabilis | |
1.262 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant F113A | Proteus mirabilis | |
1.298 | - |
1-Fluoro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant S110A | Proteus mirabilis | |
1.393 | - |
glutathione | pH 6.5, 25°C, mutant F113A | Proteus mirabilis | |
1.502 | - |
glutathione | pH 6.5, 25°C, wild-type enzyme | Proteus mirabilis | |
1.863 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, wild-type enzyme | Proteus mirabilis | |
1.95 | - |
glutathione | pH 6.5, 25°C, mutant W164A | Proteus mirabilis | |
2.155 | - |
glutathione | pH 6.5, 25°C, mutant W164A | Proteus mirabilis | |
2.432 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant S110A | Proteus mirabilis | |
3.607 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant H167A | Proteus mirabilis | |
5.509 | - |
glutathione | pH 6.5, 25°C, mutant F113A | Proteus mirabilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Proteus mirabilis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain XL1-Blue by anion exchange chromatography, dialysis, ultrafiltration, and chromatofocussing | Proteus mirabilis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.001 | - |
purified recombinant mutant W164A, pH 6.5, 25°C, substrate 4-nitrobenzyl chloride | Proteus mirabilis |
0.001 | - |
purified recombinant wild-type enzyme, pH 6.5, 25°C, substrate 4-nitrobenzyl chloride | Proteus mirabilis |
0.002 | - |
purified recombinant mutant H167A, pH 6.5, 25°C, substrate 4-nitrobenzyl chloride | Proteus mirabilis |
0.004 | - |
purified recombinant mutant S110a, pH 6.5, 25°C, substrate 4-nitrobenzyl chloride | Proteus mirabilis |
0.005 | - |
purified recombinant mutant F113A, pH 6.5, 25°C, substrate 4-nitrobenzyl chloride | Proteus mirabilis |
0.011 | - |
purified recombinant mutant F113A, pH 6.5, 25°C, substrate ethacrynic acid | Proteus mirabilis |
0.017 | - |
purified recombinant mutant H167A, pH 6.5, 25°C, substrate ethacrynic acid | Proteus mirabilis |
0.035 | - |
purified recombinant mutant S110a, pH 6.5, 25°C, substrate ethacrynic acid | Proteus mirabilis |
0.081 | - |
purified recombinant wild-type enzyme, pH 6.5, 25°C, substrate ethacrynic acid | Proteus mirabilis |
0.088 | - |
purified recombinant mutant W164A, pH 6.5, 25°C, substrate ethacrynic acid | Proteus mirabilis |
0.17 | - |
purified recombinant mutant F113A, pH 6.5, 25°C, substrate 1-chloro-2,4-dinitrobenzene | Proteus mirabilis |
0.48 | - |
purified recombinant mutant H167A, pH 6.5, 25°C, substrate 1-chloro-2,4-dinitrobenzene | Proteus mirabilis |
0.67 | - |
purified recombinant mutant F113A, pH 6.5, 25°C, substrate 1-fluoro-2,4-dinitrobenzene | Proteus mirabilis |
1.08 | - |
purified recombinant wild-type enzyme, pH 6.5, 25°C, substrate 1-chloro-2,4-dinitrobenzene | Proteus mirabilis |
1.31 | - |
purified recombinant mutant S110a, pH 6.5, 25°C, substrate 1-chloro-2,4-dinitrobenzene | Proteus mirabilis |
1.91 | - |
purified recombinant mutant H167A, pH 6.5, 25°C, substrate 1-fluoro-2,4-dinitrobenzene | Proteus mirabilis |
3.31 | - |
purified recombinant mutant S110a, pH 6.5, 25°C, substrate 1-fluoro-2,4-dinitrobenzene | Proteus mirabilis |
3.69 | - |
purified recombinant wild-type enzyme, pH 6.5, 25°C, substrate 1-fluoro-2,4-dinitrobenzene | Proteus mirabilis |
7.89 | - |
purified recombinant mutant W164A, pH 6.5, 25°C, substrate 1-fluoro-2,4-dinitrobenzene | Proteus mirabilis |
22.69 | - |
purified recombinant mutant W164A, pH 6.5, 25°C, substrate 1-chloro-2,4-dinitrobenzene | Proteus mirabilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-chloro-2,4-dinitrobenzene + glutathione | 1-chloro-2,4-dinitrobenzene is conjugated through a classic SNAr reaction that proceeds in two steps with the formation of an intermediate Meisenheimer complex | Proteus mirabilis | S-(2,4-dinitrophenyl)glutathione + HCl | - |
? | |
1-fluoro-2,4-dinitrobenzene + glutathione | 1-fluoro-2,4-dinitrobenzene is conjugated through a classic SNAr reaction that proceeds in two steps with the formation of an intermediate Meisenheimer complex | Proteus mirabilis | S-(2,4-dinitrophenyl)glutathione + HF | - |
? | |
4-nitrobenzyl chloride + glutathione | 4-nitrobenzyl chloride is conjugated through a single-step bimolecular nucleophilic substitution (SN2) with a pentacoordinate intermediate | Proteus mirabilis | S-(4-nitrobenzyl)glutathione + HCl | - |
? | |
ethacrynic acid + glutathione | ethacrynic acid is conjugated via a Michael addition reaction through the formation of an enole intermediate | Proteus mirabilis | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
GSH transferase | - |
Proteus mirabilis |
PmGST | - |
Proteus mirabilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
mutant F113A | Proteus mirabilis |
50 | - |
wild-type GST, and mutants S110A and W164A | Proteus mirabilis |
60 | - |
mutant H167A | Proteus mirabilis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.12 | - |
glutathione | pH 6.5, 25°C, mutant F113A | Proteus mirabilis | |
0.2 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant F113A | Proteus mirabilis | |
0.43 | - |
glutathione | pH 6.5, 25°C, mutant H167A | Proteus mirabilis | |
0.88 | - |
glutathione | pH 6.5, 25°C, wild-type enzyme | Proteus mirabilis | |
1.03 | - |
1-Fluoro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant F113A | Proteus mirabilis | |
1.56 | - |
glutathione | pH 6.5, 25°C, mutant S110A | Proteus mirabilis | |
2.13 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, wild-type enzyme | Proteus mirabilis | |
2.59 | - |
1-Fluoro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant H167A | Proteus mirabilis | |
3.18 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant H167A | Proteus mirabilis | |
4.37 | - |
glutathione | pH 6.5, 25°C, mutant H167A | Proteus mirabilis | |
4.7 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant S110A | Proteus mirabilis | |
6.38 | - |
glutathione | pH 6.5, 25°C, mutant F113A | Proteus mirabilis | |
6.44 | - |
glutathione | pH 6.5, 25°C, mutant S110A | Proteus mirabilis | |
6.78 | - |
1-Fluoro-2,4-dinitrobenzene | pH 6.5, 25°C, wild-type enzyme | Proteus mirabilis | |
8.66 | - |
1-Fluoro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant S110A | Proteus mirabilis | |
12.2 | - |
1-Fluoro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant W164A | Proteus mirabilis | |
12.4 | - |
glutathione | pH 6.5, 25°C, wild-type enzyme | Proteus mirabilis | |
14.99 | - |
glutathione | pH 6.5, 25°C, mutant W164A | Proteus mirabilis | |
21.44 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, mutant W164A | Proteus mirabilis | |
29.7 | - |
glutathione | pH 6.5, 25°C, mutant W164A | Proteus mirabilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
assay at | Proteus mirabilis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 9 | activity range, profiles of wild-type and mutant enzymes, overview | Proteus mirabilis |
General Information | Comment | Organism |
---|---|---|
additional information | contribution of different residues in the GSH-binding site to GSH activation, structure analysis of wild-type and mutant enzymes by circular dichroism and fluorospectroscopic analysis, overview | Proteus mirabilis |